Live Cell Imaging of T Cell Pyroptosis.
CARD8
Inflammasome
Live cell imaging
Microscopy
Pyroptosis
T cell
Journal
Methods in molecular biology (Clifton, N.J.)
ISSN: 1940-6029
Titre abrégé: Methods Mol Biol
Pays: United States
ID NLM: 9214969
Informations de publication
Date de publication:
2022
2022
Historique:
entrez:
27
6
2022
pubmed:
28
6
2022
medline:
30
6
2022
Statut:
ppublish
Résumé
Pyroptosis is a lytic, pro-inflammatory cell death program that is tightly regulated by inflammasomes in most cases. Inflammasome-dependent pyroptosis is best characterized in myeloid cells, but there is growing evidence that this cell death program also functions in human T cells. Several studies have suggested a role for inflammasome components in T-cell biology but often do not unambiguously clarify whether this means that T cells progress to pyroptosis. Pyroptosis has distinct morphological features, such as early loss of membrane integrity and ballooning, that allow it to be distinguished from apoptosis in a microscopic experiment. However, the most stringent definition of inflammasome-dependent pyroptosis is that it is genetically dependent on a pro-inflammatory caspase (caspase-1 or caspase-4) and the pore-forming protein gasdermin D (GSDMD). Therefore, using live cell imaging of T cells in combination with a genetic loss-of-function setup is the most reliable tool for us to unequivocally demonstrate that a T cell undergoes pyroptosis. Parallel live cell imaging of T cells and macrophages is limited due to the fact that T cells do not adhere while macrophages do. This can be overcome by using so-called micro-inserts that hold the cells in a limited area that can be monitored by microscopic field of view. Here we describe in detail how live cell imaging of human T cells and monocyte-derived macrophages undergoing pyroptosis can be performed.
Identifiants
pubmed: 35759205
doi: 10.1007/978-1-0716-2449-4_19
doi:
Substances chimiques
Inflammasomes
0
Intracellular Signaling Peptides and Proteins
0
Phosphate-Binding Proteins
0
Caspases
EC 3.4.22.-
Types de publication
Journal Article
Langues
eng
Sous-ensembles de citation
IM
Pagination
303-315Informations de copyright
© 2022. The Author(s), under exclusive license to Springer Science+Business Media, LLC, part of Springer Nature.
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